The recent rise of the type 1 diabetes (T1D) incidence both in Americans and globally has further increased the economic burden of the health providers worldwide. While progress has been made to understand the pathogenetic basis of T1D, we are still unable to prevent or reverse the disease. Through large-scale genetics studies in humans, others have recently identified about 50 different chromosome regions in the human genome likely containing genes that contribute to the risk or progression of T1D. Unfortunately, we still do not know for the most part which specific genes within these regions are playing a role. Further insight into the genetics of T1D will provide better risk assessment and potentially identify therapeutic targets of the disease. The Non-Obese Diabetic (NOD) mouse is of tremendous value to T1D research because of its similarity to the humans with the disease, including physiological, biochemical, and disease pathological mechanisms as well as the genetic contribution. A useful strategy to determine which genes in humans are actually playing a role is to disrupt the functions of those genes in a model like the NOD mouse. However, the current strategies for interrogating a large number of genes would take too long and are not cost effective. In the past 3 years, we have developed innovative and rapid ways of making genetically modified animal models which is highly reproducible as well as time and cost efficient. We were the first in the world to demonstrate targeted zinc-finger nuclease (ZFN) technology, and among the first to apply TAL Effector Nuclease technology, to target and disrupt ('knock out'), specific genes in the rat and have successfully applied this approach to knock out a gene in the NOD mouse. We have also developed this technology to be able to 'knock in' very specific types of gene mutations into the rat or mouse genome. These are two very important technologies which we will apply to the NOD mouse model to discover and functionally investigate a large number of the potential human T1D genes. The results of the proposed studies will have a great and broad impact on our understanding of T1D. PUBLIC HEALTH RELEVANCE: One important key to developing effective therapies for the treatment Type 1 Diabetes (T1D) is to understand the genes involved and the cells that these genes work in to target new drugs and approaches. Other studies have recently yielded important information about certain chromosome 'regions' of the human genome which likely contains one or more genes which contribute to T1D risk and/or progression. However, we do not yet know which of the many genes in these regions are important. We will use a strategy where we disrupt these potential T1D disease genes in a mouse model which develops spontaneous T1D and see what the effect of the disrupted gene is on the disease incidence and progression. We expect to uncover a significant number of new genes which are playing a role in this terrible disease and will represent new biomarkers for identifying at risk individuals and provide new targets for therapies to be developed.